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Effects of cadmium, copper, lead and zinc on growth, reproduction and survival of the earthworm Eisenia fetida (Savigny): Assessing the environmental impact of point-source metal contamination in terrestrial ecosystems.

The earthworm Eisenia fetida (Annelida: Oligochaeta) was exposed to a geometric series of concentrations of cadmium, copper, lead and zinc in artificial soil using the OECD recommended protocol. Mortality, growth and cocoon production were measured over 56 days to determine LC50 and EC50 values. No observed effect concentrations (NOECs) were also estimated. Furthermore, the percentage of viable cocoons and number of juveniles emerging per cocoon was recorded. Cocoon production was more sensitive than mortality for all the metals, particularly cadmium and copper for which NOEC reproduction values were an order of magnitude lower than those for NOEC mortality. However, there was no significant effect of metals on the viability of cocoons. The weights of earthworms declined in all treatments (including the controls) during the experiment. This was probably due to the lack of suitable food in the OECD standard soil medium used. It was concluded that future experiments should include animal manure in the test medium. The LC50, EC50 and NOEC values determined in this study were compared with concentrations of metals in soils in the vicinity of a smelting works at Avonmouth, southwest England. The 14-day LC50 for zinc in Eisenia fetida was exceeded in soils covering an area of 75 km2 around the works, compared to 4.2 km2 for copper and 4.7 km2 for lead. Soil values for cadmium did not exceed the LC50 value anywhere in the region. Similar estimates of relative effects on reproduction confirmed that zinc is most likely to be responsible for the absence of earthworms from sites close to the Avonmouth works. However, the OECD standard test overestimated the potential effects of metals on populations, since earthworms can be found as close as 1 km from the smelting works. The discrepancy between test and field observations was probably due to the greater availability of the metals in the artificial soil.

Journal Article↗

Nitrite biotransformation by mitochondria from the earthworm Eisenia foetida (Savigny).

1. Nitrite oxidase and cytochrome-c oxidase activity catalysed by cytochrome-aa3 were assayed in earthworms and rats. 2. Cytochrome-aa3 and intact mitochondria from the two species were anaerobically incubated in the presence of nitrite; the occurrence of mitochondria-induced nitrite biotransformations was evaluated by monitoring nitrite recovery in incubation medium. Possible nitric oxide production was also tested. 3. The ratio nitrite oxidase/cytochrome-c oxidase activity was much higher in earthworms than in rats. 4. Under anaerobic conditions and in the presence of respiratory substrates, earthworm mitochondria produced a time-dependent loss of nitrite in the incubation medium. On the contrary, rat mitochondria are unable to decrease environmental nitrite concentration. 5. Results support the notion that metabolic properties of earthworm mitochondria can be considered as an adaptation to chronic nitrite exposure, this toxicant being typically present in natural habitats of these worms.

Adaptation, Physiological↗

Comparative toxicity of ten organic chemicals to four earthworm species.

Ten organic chemicals were tested for toxicity to four earthworm species: Allolobophora tuberculata, Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus, using the European Economic Community's (EEC) earthworm artificial soil and contact testing procedure. The phenols were the most toxic chemicals tested, followed by the amine, substituted benzenes, halogenated aliphatic hydrocarbon, polycyclic aromatic hydrocarbon and phthalate as the least toxic chemical tested. Correlations among species within each type of test for a given chemical were extremely high, suggesting that the selection of earthworm test species does not markedly affect the assessment of a chemical's toxicity. The correlation between the two tests was low for all test species. The contact test LC50 for a given chemical cannot be directly correlated to an artificial soil test LC50 for the same earthworm species.

Amines↗

Vermiconversion of paper waste by earthworm born and grown in the waste-fed reactors compared to the pioneers raised to adulthood on cowdung feed.

The performance of four species of earthworm--Eudrilus eugeniae, Kinberg, Drawida willsi Michaelsen, Lampito mauritii, Kinberg and Perionyx excavatus, Perrier--born and grown in vermireactors fed with paper waste was studied over six months, in terms of vermicast output per unit feed, production of offspring, and increase in worm zoomass. These were compared with the performance of the previous generation which had been raised to adulthood on cowdung as principal feed before shifting them to vermireactors operating on cowdung-spiked paper waste. The results indicated that except with D. willsi of which the second generation performed only a shade better than the first, there was significant improvement in vermicast output, animal growth, and reproduction in the second generation compared to the first. The results indicated that cowdung-spiked paper waste can be an adequate food for successive generations of earthworms and that reactors can be operated indefinitely on this feed. The results also indicated that the earthworm generations born and raised in vermireactors operated on this feed become better vermiconverters of this feed than the parent earthworms.

Animals↗

Vermistabilization of textile mill sludge spiked with poultry droppings by an epigeic earthworm Eisenia foetida.

Investigations were made to explore the potential of an epigeic earthworm Eisenia foetida to transform textile mill sludge spiked with poultry droppings in to value added product, i.e., vermicompost. The growth and reproduction of E. foetida was monitored in a range of different feed mixtures for 77 days in the laboratory under controlled experimental conditions. The maximum growth was recorded in 100% cow dung (CD). Replacement of poultry droppings by cow dung in feed mixtures and vice versa had little or no effect on worm growth rate and reproduction potential. Worms grew and reproduced favourably in 70% poultry droppings (PD)+30% solid textile mill sludge (STMS) and 60% PD+40% STMS feed mixtures. Greater percentage of STMS in the feed mixture significantly affected the biomass gain and cocoon production. Net weight gain by earthworms in 100% CD was 2.9-18.2 fold higher than different STMS containing feed mixtures. The mean number of cocoon production was between 23.4+/-4.65 (in 100% CD) and 3.6+/-1.04 (in 50% PD+50% STMS) cocoons earthworm(-1) for different feed mixtures tested. Vermicomposting resulted in significant reduction in C:N ratio and increase in nitrogen and phosphorus contents. Total potassium, total calcium and heavy metals (Fe, Zn, Pb and Cd) contents were lower in the final product than initial feed mixtures. Our trials demonstrated vermicomposting as an alternate technology for the recycling and environmentally safe disposal/management of textile mill sludge using an epigeic earthworm E. foetida if mixed with poultry droppings.

Animals↗

A microcosm approach to assessing the effects of earthworm inoculation and oat cover cropping on CO2 fluxes and biological properties in an amended semiarid soil.

We designed a microcosm experiment to assess the influence of inoculation with Eisenia foetida earthworms and the establishment of an Avena sativa cover crop on biological (enzyme activities and labile carbon fractions) soil quality indicators in a soil treated with a composted organic residue, and to determine the contribution of these treatments to carbon dioxide emissions from the soil to the atmosphere of the microcosm. The microcosms were incubated for 53 days under 28 degrees C/18 degrees C day/night temperatures. The addition of earthworms and the planting of A. sativa increased dehydrogenase activity of compost amended soil by about 44% after 23 days of incubation. The metabolic potential, calculated as the ratio dehydrogenase activity/water soluble C, was higher in the compost amended soil planted with A. sativa. The highest total amount of CO2-C evolved occurred in the soil treated with composted residue and earthworms (about 40% of the total amount of CO2 evolved came from earthworm activity). The planting of A. sativa increased the decomposition rate constant of organic matter in the amended soil but decreased the potentially mineralizable C pool. In conclusion, the establishment of an A. sativa cover crop and the addition of E. foetida to a degraded agricultural soil treated with composted residue were effective treatments for improving the biological and biochemical quality and the metabolic potential of the soil.

Animals↗

Effect of experimental microbial challenge on the expression of defense molecules in Eisenia foetida earthworm.

Earthworms are able to protect themselves against invading pathogens due to efficient innate defense mechanisms. Currently, two types of antimicrobial factors including lysozyme-like molecule and factors with hemolytic activity, as well as a pattern recognition protein named coelomic cytolytic factor (CCF) have been identified in Eisenia foetida earthworms. However, the modulations of these defense molecules during in vivo immune response have not been addressed. In this study, we investigated the effect of experimental challenge with live Gram-negative and Gram-positive bacteria and with beta-1,3-glucan on the expression of CCF and the hemolytic factor fetidin. In parallel, we followed levels of hemolytic activity and lysozyme-like activity in the coelomic fluid of challenged earthworms. We show that the biosynthesis of CCF, but not fetidin, is up-regulated upon microbial stimulation. Parenteral administration of bacteria or microbial polysaccharides in earthworms results, in the coelomic fluid, in augmented level of CCF, increased lysozyme-like activity and decreased hemolytic activity. The decreased hemolytic activity of the coelomic fluid reflects the increase of the whole protein content in the absence of synthesis of hemolytic proteins.

Animals↗

Modeling ingestion as an exposure route for organic chemicals in earthworms (Oligochaeta).

Earthworms take up chemicals from both soil pore water and food, but the quantitative contribution of each route is unclear. In this paper, a model is presented for the accumulation of organic chemicals in earthworms, including a compartment for the gut contents. A Monte Carlo screening method is used to calibrate the model simultaneously to four experimental data sets for hexachlorobenzene (HCB) in the compost worm (Eisenia andrei). The calibration procedure shows that the dominant route of exposure is across the gut wall. Nevertheless, predicted body residues of HCB are generally no more than 20% higher than the estimate based on equilibrium partitioning (EP), independent of the uncertainties in all of the model parameters. The deviation from EP is mainly related to the digestive efficiency of the earthworm. Body residues are not expected to exceed the EP estimate by more than 50% for other chemicals or other earthworm species. The model is particularly useful when the organism's food source is specifically contaminated and EP cannot be applied.

Administration, Oral↗

The implications of copper fungicide usage in vineyards for earthworm activity and resulting sustainable soil quality.

To investigate the impact of copper-containing fungicides (copper oxychloride) on earthworms in South African vineyards, field inventories of earthworms in and between vine rows were carried out and compared to directly adjacent grassland. Also copper content, pH, organic matter content, and soil porosity were determined in these soils. This was combined with laboratory experiments to study the impact of vineyard soil characteristics on the burrowing and dispersal behavior of earthworms. Moreover, the direct toxic action of copper oxychloride on different endpoints of the earthworms (survival and growth) was studied. Copper oxychloride had a negative impact on these endpoints (decreased growth and survival related to increased copper body content) as well as on the behavioral aspect (decreased burrowing rate and avoidance of copper-containing soil). Moreover, there was an inverse relation between burrowing activity and soil bulk density that could also be related to the copper content. This may lead to a decrease in sustainable soil quality in vineyards.

Agriculture↗

The impact of organophosphate pesticides in orchards on earthworms in the Western Cape, South Africa.

Earthworm population density was measured in and adjacent to an orchard in an agricultural area in the Western Cape, South Africa. Worm densities were very low in orchards (22/m(2)) compared to adjacent uncultivated fields (152/m(2)) at a distance from the orchards. The possible effect of organophosphate pesticides on the earthworms was investigated. Background soil concentrations of chlorpyrifos prior to the start of the spraying season were low (0.2-2.7 microg/kg) but persistent for up to 6 months after the last spraying event, and the pesticide was, as a result of rainfall, transported to nontarget areas by runoff. Background concentrations of azinphos methyl were higher than those of chlorpyrifos (1.6-9.8 microg/kg) but not detectable 2 weeks after a spraying event. Azinphos methyl was mostly transported by wind (spray drift) to adjacent areas. A microcosm study indicated effects of chlorpyrifos on earthworms as determined by measuring biomass change and Cholinesterase inhibition. It is concluded that earthworms were affected detrimentally by the pesticides due to chronic (chlorpyrifos) and intermittent (azinphos methyl) exposure.

Agriculture↗

Survival, Pb-uptake and behaviour of three species of earthworm in Pb treated soils determined using an OECD-style toxicity test and a soil avoidance test.

Mature (clitellate) Eisenia andrei Bouché (ultra epigeic), Lumbricus rubellus Hoffmeister (epigeic), and Aporrectodea caliginosa (Savigny) (endogeic) earthworms were placed in soils treated with Pb(NO(3))(2) to have concentrations in the range 1,000 to 10,000 mg Pb kg(-1). After 28 days LC50(-95%confidence limit)(+95%confidence limit) values were E. andrei 5824(-361)(+898) mg Pb kg(-1), L. rubellus 2867(-193)(+145) mg Pb kg(-1) and A. caliginosa2747(-304)(+239) mg Pb kg(-1) and EC50s for weight change were E. andrei2841(-68)(+150) mg Pb kg(-1), L. rubellus1303(-201)(+240) mg Pb kg(-1) and A. caliginosa1208(-206)(+212) mg Pb kg(-1). At any given soil Pb concentration, Pb tissue concentrations after 28 days were the same for all three earthworm species. In a soil avoidance test there was no difference between the behaviour of the different species. The lower sensitivity to Pb exhibited by E. andrei is most likely due to physiological adaptations associated with the modes of life of the earthworms, and could have serious implications for the use of this earthworm as the species of choice in standard toxicological testing.

Adaptation, Physiological↗

In situ localization and substrate specificity of earthworm protease-II and protease-III-1 from Eisenia fetida.

Recently, the function in fibrinolysis of earthworm proteases has been studied. In our experiments, earthworm protease-II (EfP-II) and earthworm protease-III-1 (EfP-III-1) were isolated and purified from Eisenia fetida. As shown by the assay of sections of the earthworm on fibrin plates, the enzymic activity was mainly detected around the clitellum. In the presence of anti-EfP-II or anti-EfP-III-1 serum, the immunological signals of the two isozymes were clearly found in the anterior alimentary mucosa, suggesting that EfP-II and -III-1 are localized and expressed in intestinal epithelial cells. The Michaelis-Menten constant (K(m)) for EfP-III-1 reacting with BAEE is smaller (1.7x10(-5)M) in comparison with the K(m) values of other substrates such as Chromozym-Try and -TH (3.3-6.0x10(-5)M). This indicates that EfP-III-1 is a trypsin-like protein. EfP-II shows a strong trypsin-like, moderate elastase-like and weak chymotrypsin-like serine function. The relative broad substrate specificity of EfP-II and EfP-III-1 is consistent with their localization in the anterior alimentary canal where different micro-organisms and ingested proteins require to be digested.

Animals↗

Modulation of motor patterns by sensory feedback during earthworm locomotion.

Role of sensory feedback to motor pattern activity concerning locomotion in the earthworm, Eisenia fetida, was investigated. We have previously reported that bath application of octopamine induces fictive locomotion in the earthworm. In this study, we have examined the role of sensory feedback during fictive locomotion by analyzing electrical activities from the cut end and intact first lateral nerves of the ventral nerve cord (VNC). From the cut end recordings, motor activity associated with fictive locomotion was measured. A mixture of sensory and motor activities was measured from the intact first lateral nerve using en passant recordings, and sensory activity was separated by subtraction of the cut end recording (mainly motor activities) from the intact first lateral nerve recording. We estimated the effect of sensory feedback from the earthworm body wall by comparing recordings that made when the preparation was in-contact with a substrate or suspended above it. Motor pattern activities and the coefficient of variation for inter-spike-interval of motor outputs were increased under suspended conditions during circular muscle contraction. These results indicate that sensory feedback modulates the pattern of motor activity in the earthworm during locomotion.

Action Potentials↗

Biological assessment of contaminated land using earthworm biomarkers in support of chemical analysis.

Biological indicators can be used to assess polluted sites but their success depends on the availability of suitable assays. The aim of this study was to investigate the performance of two earthworm biomarkers, lysosomal membrane stability measured using the neutral red retention assay (NRR-T) and the total immune activity (TIA) assay, that have previously been established as responsive to chemical exposure. Responses of the two assays were measured following in situ exposure to complexly contaminated field soils at three industrial sites as well as urban and rural controls. The industrial sites were contaminated with a range of metal (cadmium, copper, lead, zinc, nickel and cobalt) and organic (including polycyclic aromatic hydrocarbons) contaminants, but at concentrations below the 'New Dutch List' Intervention concentrations. Exposed earthworms accumulated both metals and organic compounds at the contaminated sites, indicating that there was significant exposure. No effect on earthworm survival was found at any of the sites. Biomarker measurements, however, indicated significant effects, with lower NRR-T and TIA found in the contaminated soils when compared to the two controls. The results demonstrate that a comparison of soil pollutant concentrations with guideline values would not have unequivocally identified chemical exposure and toxic effect for soil organisms living in these soils. However, the earthworm biomarkers successfully identified significant exposure and biological effects caused by the mixture of chemicals present.

Animals↗

Lead bioaccumulation in earthworms, Lumbricus terrestris, from exposure to lead compounds of differing solubility.

This study examined the relative effects of soluble and less soluble Pb compounds on Pb accumulation by Lumbricus terrestris. The earthworms were exposed to composted cattle manure contaminated with a range of concentrations of either soluble lead acetate trihydrate (PbAc) (14.5, 72.2, 137, 257, and 603 microg/g) or less soluble lead carbonate (Pb(CO(3))(2)) (5.09, 171, 575, and 710 microg/g). Relative Pb bioaccumulation rates in earthworms from the PbAc and Pb(CO(3))(2) lead carbonate trials were compared over time (0, 20, 40, and 60 days). The two molecular forms of Pb were heavily adsorbed onto the organic matter. Relative soluble Pb concentrations in the manure pore water were very low, at 0.049% and 0.022% of total Pb for PbAc and Pb(CO(3))(2), respectively. There was a significant uptake of lead by earthworms from both the readily soluble and less soluble forms of Pb. Regression surfaces relating organic matter total Pb concentration to exposure time and Pb body burden were compared. For the PbAc trial, the following equation was fitted: Body burden=2.3429+0.0014 x--0.0632 y+0.0008 xy (R(2)=0.8494, p<0.0001). For the Pb(CO(3))(2) trial, the equation was: Body burden=-2.0760--0.0067 x-0.2297 y-0.0038 y(2)+0.0003 xy (R(2)=0.5686, p<0.0001) (for x=organic matter Pb concentration in microg/g and y=time in days). The Pb bioaccumulation resulting from the PbAc exposure was significantly higher than that from Pb(CO(3))(2). Body burdens of earthworms exposed to PbAc were linearly related to exposure time and total Pb concentration. Worms exposed to Pb(CO(3))(2) had non-linear Pb accumulation over time that suggested a decrease in Pb bioavailability with increasing exposure time. The results indicate that L. terrestris feeding on detritus in the upper soil levels can mobilize and accumulate Pb from the principal transformation products of metallic Pb, as at shooting ranges and lead smelter sites.

Animals↗

Comparative distribution of urocortin- and CRF-like immunoreactivities in the nervous system of the earthworm Lumbricus terrestris.

Corticotropin-releasing factor (CRF) and urocortin (Ucn) are both members of the CRF neuropeptide family. The distribution of Ucn- and CRF-like immunoreactive (ir) structures in the central nervous system of several vertebrate species has been studied, but little is known about that in non-vertebrates. We used a highly specific polyclonal antibody against rat Ucn and CRF to determine and compare the distribution of Ucn- and CRF-like immunoreactivity in the earthworm nervous system. Several Ucn- and CRF-like ir perikarya were described in the cerebral ganglion, subesophageal and ventral cord ganglia. The majority of Ucn-like ir cells were found in the ventral ganglia, whereas CRF-like ir cells were most abundant in the cerebral ganglion. Scattered Ucn- and CRF-like ir varicose fiber terminals were seen in all areas of the earthworm central nervous system. Ucn-like ir cell bodies and fiber terminals were also demonstrated in the pharyngeal wall. No co-localization of Ucn- and CRF-like ir nervous structures were observed. This study provided morphological evidence that Ucn- and CRF-like neurosecretory products exist in the earthworm central nervous system. Furthermore, both the distribution and morphology of Ucn- and CRF-like ir structures were distinct, therefore, it can be hypothesized that these neuropeptides exert different neurendocrine functions in the earthworm nervous system.

Animals↗

My saga with earthworms.

Earthworms have been studied as a readily available, easily maintainable and cheap test species to develop toxicity test systems as an alternative to in vivo rodent bioassays. The results obtained, mainly with metals, since the programme was initiated about 15 years ago, are summarized in this review. Metals proved more toxic to rodents than earthworms; the relative order of toxicity for different metals was similar in both species. The effects of metals and other compounds on enzyme activity [glutathione S-transferase (GST) and catalase] and the induction of metallothioneins in earthworms are described. It is concluded that useful preliminary information can be obtained in earthworms on the acute toxicity of inorganic metal ions and on the relative order of toxicity of groups of metal salts.

Animals↗

Influence of nematophagous fungi, earthworms and dung burial on development of the free-living stages of Ostertagia (Teladorsagia) circumcincta in New Zealand.

Biological options for nematode parasite control are being sought, as the long-term efficacy of conventional anthelmintics comes increasingly under threat from drug-resistant parasites. Three biological methods with the potential to reduce pasture contamination by parasitic nematode larvae were examined: (a) killing of larvae developing in dung by nematophagous fungi; (b) removal of dung through earthworm ingestion; (c) burial of dung in soil as might occur through the action of dung beetles. Field trials with the test bio-control agents were carried out in autumn and spring by adding dung from sheep infected with Ostertagia (Teladorsagia) circumcincta to pots of ryegrass/white clover. The factorial treatment structure included five fungal treatments (individual applications of Duddingtonia flagrans, Monacrosporium gephyropagum and Harposporium helicoides, a combination of all the three fungi together and an untreated control), two dung burial treatments (dung buried or deposited on the soil surface) and two earthworm treatments (earthworms present or absent). D. flagrans and H. helicoides, individually or in combination, reduced recovery of infective stage larvae in experiment 1, while only H. helicoides reduced recovery in experiment 2. In both the experiments, dung burial increased the total number of larvae recovered, while the number of infective larvae were reduced by the action of earthworms. Increased recovery following burial, along with the fact that larvae moved rapidly from soil onto herbage, suggests that soil may provide a protective reservoir for infective larvae infesting herbage.

Animals↗